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Secretomes of mesenchymal stromal cells and leukocyte and platelet rich fibrin

ABSTRACT: The objective of the present study was to compare the secretome of BMSC (BMSC-CM) to that of L-PRF (LPRF-CM) – an established standard for GF therapy, in the context of wound healing and regeneration.

INSTRUMENT(S): LTQ Orbitrap Elite

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Stem Cell, Cell Culture

DISEASE(S): Wounds And Injuries

SUBMITTER: Siddharth Shanbhag

LAB HEAD: Siddharth Shanbhag

PROVIDER: PXD041617 | Pride | 2023-10-24

REPOSITORIES: Pride

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			Action	DRS
	BM-1.mzML	Mzml
	BM-1.raw	Raw
	BM-4.raw	Raw
	BM-9.raw	Raw
	BM-P.raw	Raw

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1 - 5 of 11

Publications

Proteomic Analysis of Mesenchymal Stromal Cells Secretome in Comparison to Leukocyte- and Platelet-Rich Fibrin.

Al-Sharabi Niyaz N Gruber Reinhard R Sanz Mariano M Mohamed-Ahmed Samih S Kristoffersen Einar K EK Mustafa Kamal K Shanbhag Siddharth S

International journal of molecular sciences 20230822 17

Secretomes of mesenchymal stromal cells (MSCs) are emerging as a novel growth factor (GF)-based strategy for periodontal and bone regeneration. The objective of this study was to compare the secretome of human bone marrow MSC (BMSC) to that of leukocyte- and platelet-rich fibrin (L-PRF), an established GF-based therapy, in the context of wound healing and regeneration. Conditioned media from human BMSCs (BMSC-CM) and L-PRF (LPRF-CM) were subjected to quantitative proteomic analysis using liquid ...[more]

PMID: 37685865

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Deciphering the protein secretome of leukocyte-platelet rich fibrin

Project description:Leukocyte-platelet rich fibrin (L-PRF) is extensively used in the dentistry field and other clinical scenarios due to its regeneration properties. We obtained L-PRF membranes in absence of anticoagulants and cultured in DMEM. The secretomewas collected at days 3, 7 and 21 in order to identify the proteins secreted and the differences over time. Protein secretomeat day 3 was analysed by LC-MS/MS and differences in the proteome were analysed by Sequential Window Acquisition of all Theoretical Mass Spectra (SWATH). Overall, 705 proteins were identified in the secretome of L-PRF membranes after 3 days of culture, including growth factors (EGF, PDGFA) and proteins related to platelet and neutrophil degranulation. A total of 202 differentially secreted proteins were quantified by SWATH when comparing secretomes at days 3, 7 and 21. The majority of them were enriched at day 3 such as MMP9, TSP1 and CO3. On the contrary, fibrinogen and CATS were found down-regulated at day 3. Growth factor and western blotting analysis corroborated the proteomic results. This is the most detailed proteome analysis of the L-PRF secretome to date. Proteins and growth factors identified, and their kinetics, provide novel information to further understand the wound healing properties of L-PRF.

2021-09-09 | PXD017963 | Pride

TGFβ activity released from platelet-rich fibrin adsorbs to titanium surface and collagen membranes

Project description:Considering that PRF is clinically used in combination with dental implants and collagen membranes, we exposed titanium discs and collagen membranes to PRF lysates. We showed here that PRF-derived TGF-β activity adsorbs to titanium implants and collagen membranes indicated by the changes in gene expression and immunoassay analysis. Our study points towards TGF-β as major target of PRF and suggest that TGF-β activity released by PRF adsorbs to titanium surface and collagen membranes.

2020-06-24 | PXD014382 | Pride

Expression data from hepatitis E virus inoculated PLC/PRF/5 cells

Project description:Hepatitis E virus (HEV) is an important causative pathogen of acute hepatitis. Because of the absence of an in vitro culture system for HEV, research has been greatly impeded. And interaction between HEV and host cells was mainly studied by tansfection/transinfection system, such as Adeno virus transinfection system. We developed an in vitro culture system for HEV in PLC/PRF/5 cells. With this in vitro culture system, we studied the gene expression profile change by HEV infection. Using a microarray assay, we analysed genes of PLC/PRF/5 cells, whose transcription level could be changed by HEV infection. Five flasks of PLC/PRF/5 cells inoculated with HEV were used as test sample, and five flasks inoculated with serum-free DMEM/199 medium were used as control samples. Both test and control flasks were cultured under the same conditions. Sixty days after inoculation, the test and control flasks was resolved with Trizol and analysed with Affymetrix HG-U133 Plus 2 array.

2014-01-01 | E-GEOD-53731 | biostudies-arrayexpress

Expression data comparing chemoresistant (cisplatin and doxorubicin) hepatospheres and differentiated HCC cells

Project description:To identify candidate genes that are up-regulated in chemoresistant (cisplatin and doxorubicin) hepatospheres as compared with their differentiated counterparts Affymetrix Human Genome U133 Plus GeneChip 2.0 PLC/PRF/5 HCC cells were grown as spheroids and treated with cisplatin and doxorubicin combination. Differentiated counterparts were generated by addition of FBS in the spheroids and allowed to attach.

2014-06-01 | E-GEOD-53005 | biostudies-arrayexpress

Human gingival fibroblasts: control vs. lysophosphatidic acid (LPA 18:1)-treated

Project description:Transcriptional profiling of pass 2 primary human gingival fibroblasts (GF) comparing control untreated GF with GF treated with LPA 18:1 for 2h or 8h. The goal of this study was to determine the effects of LPA 18:1 on global GF gene expression. Three-condition experiment, GF vs. LPA-treated (2h, 8h) GF cells. Biological replicates: 3 control replicates, 3 LPA-treated replicates.

2014-05-12 | E-GEOD-57496 | biostudies-arrayexpress

Gene expression analyses of HUVECs treated with conditioned medium derived from bone marrow mesenchymal stromal cells (BMSC-CM)

Project description:A functional vascular system is a prerequisite for bone repair as disturbed angiogenesis often causes non-union. Paracrine factors released from human bone marrow mesenchymal stromal cells (BMSCs) have angiogenic effects on endothelial cells. However, whether these paracrine factors participate in blood flow dynamics within bone capillaries remains poorly understood. Here, we used two different microfluidic designs to investigate critical steps during angiogenesis and found pronounced effects of endothelial cell proliferation as well as chemotactic and mechanotactic migration induced by BMSC conditioned medium (CM). The application of BMSC-CM in dynamic cultures demonstrates that bioactive factors in combination with fluidic flow-induced biomechanical signals significantly enhanced endothelial cell migration. Transcriptional analyses of endothelial cells demonstrate the induction of a unique gene expression profile related to tricarboxylic acid cycle and energy metabolism by the combination of BMSC-CM factors and shear stress, which opens an interesting new avenue to explore during fracture healing. Our results stress the importance of in vivo - like microenvironments simultaneously including biochemical, biomechanical and oxygen levels when investigating key events during vessel repair.

2023-01-01 | GSE208335 | GEO

Angiogenic Properties of 'Leukocyte- and Platelet-Rich Fibrin'.

Project description:Leukocyte- and Platelet-Rich Fibrin (L-PRF) is an autologous platelet concentrate, consisting of a fibrin matrix enriched with platelets, leukocytes and a plethora of cytokines and growth factors. Since L-PRF is produced bedside from whole blood without the use of an anti-coagulant, it is becoming a popular adjuvant in regenerative medicine. While other types of platelet concentrates have been described to stimulate blood vessel formation, little is known about the angiogenic capacities of L-PRF. Therefore, this study aimed to fully characterize the angiogenic potential of L-PRF. With an antibody array, the growth factors released by L-PRF were determined and high levels of CXC chemokine receptor 2 (CXCR-2) ligands and epidermal growth factor (EGF) were found. L-PRF induced in vitro key steps of the angiogenic process: endothelial proliferation, migration and tube formation. In addition, we could clearly demonstrate that L-PRF is able to induce blood vessel formation in vivo, the chorioallantoic membrane assay. In conclusion, we could demonstrate the angiogenic capacity of L-PRF both in vitro and in vivo, underlying the clinical potential of this easy-to-use platelet concentrate.

| S-EPMC6168453 | biostudies-literature

Slu7 is essential for liver differentiation, metabolism and quiescence

Project description:The equilibrium between cellular differentiation and proliferation is fundamental for tissue homeostasis. This is particularly important for the liver, a highly differentiated organ with systemic metabolic functions still endowed with unparalleled regenerative potential. Hepatocellular de-differentiation and uncontrolled proliferation are at the basis of liver carcinogenesis. We have identified SLU7, a pre-mRNA splicing regulator inhibited in hepatocarcinoma as a pivotal gene for hepatocellular homeostasis. SLU7 knockdown in human liver cells and mouse liver resulted in profound changes in pre-mRNA splicing and gene expression, leading to impaired glucose and lipid metabolism, refractoriness to key metabolic hormones, and reversion to a fetal-like gene expression pattern. Hepatocellular proliferation and a switch to a tumor-like glycolytic phenotype were also observed. Mechanistically, SLU7 governed the splicing and/or expression of essential genes for hepatocellular differentiation like SRSF3 and HNF4a, and was identified as a critical factor in cAMP-regulated gene transcription. SLU7 is therefore central for hepatocyte identity and quiescence. The expression of the splicing factor SLU7 in the human hepatocellular carcinoma cell line PLC/PRF/5 was knocked down with specific siRNAs (siSlu7). An irrelevant siRNA (siGL2) was used as negative control. The RNAto perfrom the array analysis was extracted 48h after transfection.

2014-05-29 | E-GEOD-54090 | biostudies-arrayexpress

Anti-VLDL receptor monoclonal antibodies inhibit fibrin-VLDL receptor interaction and reduce fibrin-dependent leukocyte transmigration.

Project description:Our previous studies revealed that the interaction of fibrin with the very low density lipoprotein receptor (VLDLR) promotes transendothelial migration of leukocytes and thereby inflammation, and localised the fibrin-binding site to CR-domains 2-4 of this receptor. In the present study, we tested interaction of three anti-VLDLR monoclonal antibodies, mAb 1H10, 1H5, and 5F3, with recombinant fragments of VLDLR containing various combinations of its CR-domains and found that the epitopes for mAb 1H10 and mAb 1H5 overlap with the fibrin-binding site of VLDLR. Based on these findings, we hypothesised that mAb 1H10 and mAb 1H5 should inhibit fibrin-VLDLR interaction and modulate leukocyte transmigration. To test this hypothesis, we first demonstrated that these monoclonal antibodies both have high affinity to the fibrin-binding fragments of the VLDL receptor and efficiently inhibit interaction between the VLDLR-binding fragment of fibrin and the fibrin-binding fragments of VLDLR. Next, in the in vitro experiments using leukocyte transendothelial migration assay we found that both monoclonal antibodies efficiently inhibit leukocyte transmigration induced by fibrin mimetic NDSK-II. Finally, in vivo experiments using mouse model of peritonitis revealed that mAb 1H10 and mAb 1H5 both significantly reduce infiltration of leukocytes into the peritoneum. Furthermore, our experiments using mouse model of myocardial ischemia-reperfusion injury revealed that both monoclonal antibodies significantly reduce myocardial injury induced by ischaemia-reperfusion. Thus, the results obtained indicate that monoclonal antibodies 1H10 and 1H5 are novel specific inhibitors of fibrin-VLDLR-dependent leukocyte transmigration pathway. They may represent potential therapeutics for treatment of fibrin-dependent inflammation including myocardial ischaemia-reperfusion injury.

| S-EPMC5294920 | biostudies-literature

RNAseq of gingival fibroblasts exposed to PRF membranes and PRF serum.

Project description:Platelet-rich fibrin (PRF) is prepared from the coagulated plasma of fractionated blood. When squeezing between two plates, PRF is separated into the solid PRF membranes and a liquid exudate, the PRF serum. The question arises regarding the extent to which the overall PRF activity remains in the membranes and what is lost in the serum. To this aim, we have exposed gingival fibroblasts to lysates prepared from PRF membranes and PRF serum, followed by bulk RNA sequencing. A total of 268 up- and 136 down-regulated genes in gingival fibroblasts exposed to PRF lysates are significantly regulated under the premise of a minimum log2 2.5-fold change and a minus log10 significance level of two, respectively. PRF serum caused 62 up- and 32 down-regulated genes when gingival fibroblasts were exposed to PRF serum, respectively. Among the 61 genes commonly up-regulated by PRF lysate and serum were CXCL1, CXCL5, CXCL6, CXCL8, IL33, and IL6 and PTGS2, STC1. PRF lysate further increased the chemokines CCL2, CCL7, CXCL2, CXCL3, and the IL1R1, IL1RL1, and IL1RN – as well as the paracrine factors IL11, LIF, IGF1, BMP2, BMP6, FGF2, CCN2/CTGF and HAS1, HAS2, HAS3. The 16 up-regulated genes by PRF serum included DKK1. Among the 122 down-regulated genes by PRF lysate were IFIT1, IFIT2, IFIT3, OSR1, OSR2. Among the 32 down-regulated genes by PRF serum were FGF18 and GDF15. Taken together, PRF lysates, compared to PRF serum, cause a more complex response of gingival fibroblasts with a chemokine with an obvious increase in chemokine expression and spectrum of paracrine factors.

2024-05-31 | GSE268248 | GEO

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